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PQDx 0106-038-00 WHO PQDx PR July/2015, version 2.0

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WHO Prequalification of In Vitro Diagnostics PUBLIC REPORT

Product: DS-EIA-HIV-AGAB-SCREEN

Number: PQDx 0106-038-00

Abstract DS-EIA-HIV-AGAB-SCREEN with product codes I-1654, I-1652 and I-1656, manufactured RPC Diagnostic Systems, CE-mark regulatory version, was accepted for the WHO list of prequalified in vitro diagnostics and was listed on 13 July 2015. DS-EIA-HIV-AGAB-SCREEN kit is a fourth generation immunoassay utilizing a mixture of antigens and antibodies for the qualitative simultaneous detection IgG, IgM and IgA antibodies to all known subtypes of HIV-1, HIV-2, HIV-1 group O and HIV-1 p24 antigen at the early stage of infection in human serum or plasma. The kit is intended for the in vitro diagnostic screening. The kit is for in vitro diagnostic use by healthcare professionals and will not be sold to the general public. The sensitivity of the kit is 0.5 IU/ml (HIV-1 p24 ANTIGEN 1st international reference reagent, NIBSC Code: 90/636) when HIV-1 p24 antigen detecting. DS-EIA-HIV-AGAB-SCREEN is a one-step ”sandwich” assay based on microwells coated by a recombinant antigens representing an immunodominant regions of HIV-1, HIV-1 (O), HIV-2 proteins and antibodies raised against p24 of HIV-1. The Conjugate is a mixture of the other viral antigen epitopes all labelled with biotin and horseradish peroxidase, and different antibodies, also raised against p24, labelled with biotin. Serum or plasma specimens are added to these wells and if p24 antigens and/or antibodies specific for HIV-1, HIV-1 (O) and HIV-2 are present in the specimen, they form stable complexes with the HIV antigens or antibodies attached to the well. Antigen/antibody complexes are then identified by the addition of: (1) biotinylated antigens and antibodies and (2) horseradish peroxidase (HRP) streptavidin conjugate and HRP labelled antigens. Unbound conjugates are removed by washing. After adding of a solution containing 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide wells with bound Conjugate develop a blue colour which is converted into yellow colour which may be read at 450nm or 450/620nm after the reaction has been stopped with sulphuric acid. The test kit contains:

Product code I-1654

Product code I-1652

Product code I-1656


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HIV AG/AB Coated Strips Polystyrene stripped 96-well (breakable wells) plate coated with mixture of purified recombinant antigens – gp160, p31 HIV-1, gp41 HIV-1 group O, gp36 HIV-2, and antibodies to antigen p24 HIV-1.

96 tests/ 1 plate

192 tests/ 2 plates

480 tests/ 5 plates

Conjugate-1 (concentrated 11-fold) Mixture of biotinylated recombinant antigens gp160, p31 HIV-1, gp41 group O, gp36 HIV-2, and antibodies to antigen p24 HIV-1. Preserving agent 0.01% gentamycin sulfate, 0.01% thimerosal.

1 vial of 1.2 ml

1 vial of 1.2 ml

1 vial of 1.2 ml or 3 vials of 3.6 ml

Conjugate-2 (concentrated 11-fold) Mixture of horseradish peroxidase labelled recombinant antigens gp41, gp36 HIV-2 and horseradish peroxidase labelled streptavidin. Preserving agent 0.02% phenol.

1 vial of 2.0 ml

1 vial of 2.0 ml

2 vials of 2.0 ml or 1 vial of 4ml

Conjugate-1 diluent Transparent or slightly opalescent liquid, orange-coloured. Preserving agent 0.01% gentamycin sulfate, 0.02% phenol.

1 vial of 12.0ml

1 vial of 12.0ml

3 vials of 12.0ml

Conjugate-2 diluent Transparent or slightly opalescent liquid, blue-coloured. Preserving agent 0.01% thimerosal.

1 vial of 20ml

1 vial of 20ml

2 vials of 20ml

AB Positive Control, Inactivated Heat inactivated human plasma positive for anti-HIV, negative for antigen p24 HIV-1, HBsAg and anti-HCV. Transparent or slightly opalescent liquid, orange-coloured. Preserving agent 0.01% gentamycin sulfate, 0.025% thimerosal.

1 vial of 2.0ml

1 vial of 2.0ml

1 vial of 4.0ml or 2 vials of 2.0 ml

AG Positive Control, Inactivated Purified recombinant antigen p24 HIV-1 in heat inactivated human plasma negative for anti-HIV, anti-HCV, HBsAg and p24 HIV-1 antigen. Transparent or slightly opalescent liquid, red-coloured. Preserving agent 0.01% gentamycin sulfate, 0.05% thimerosal.

1 vial of 2.0ml

1 vial of 2.0ml

1 vial of 4.0ml or 2 vials of 2.0 ml

Negative Control, Inactivated Heat inactivated human plasma negative for anti-HIV, negative for antigen p24 HIV-1, HBsAg and anti-HCV. Transparent or slightly opaque liquid, green-coloured. Preserving agent 0.01% thimerosal.

1 vial of 2.5ml

1 vial of 2.5ml

2 vials of 2.5ml or 1 vial of 5.0 ml

Washing Solution (concentrated 25-fold) Transparent or slightly opaque liquid, colourless or

1 vial of 50.0ml

1 vial of 120.0ml

2 vials of 120.0ml


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pale yellow.

Substrate buffer Citric acid and sodium acetate solution containing H2O2. Transparent colourless liquid. Preserving agent 0.04% ProClin 300.

1 vial of 25.0ml

1 vial of 25.0ml

3 vials of 25.0ml or 2 vials of 50.0ml

TMB Solution containing Tetramethyl-benzidine (TMB).

1 vial of 2.5ml

1 vial of 2.5ml

3 vials of 2.5 ml or 2 vials of 3.5ml

Stopping Reagent 0.2M sulphuric acid solution. Transparent colourless liquid.

1 vial of 25.0ml

2 vials of 25.0ml or 1 vial of 50.0ml

4 vials of 25.0 ml or 2 vials of 50.0ml

Protective film for EIA plates 2 pcs 4 pcs 10 pcs

Plastic clip or polyethylene bag with a zip lock 1 pc 2 pcs 3 pcs

Plastic dish for liquid reagents 2 pcs 4 pcs 10 pcs

Disposable tips 16 pcs 32 pcs 80 pcs

Instructions for use 1 pc 1 pc 1 pc

Storage: The test kit should be stored at 2 - 8 °C. Shelf-life: 24 months.

Summary of prequalification status for DS-EIA-HIV-AGAB-SCREEN

Initial acceptance

Date Outcome

Status on PQ list 13 July 2015 listed

Dossier assessment 02 March 2015 MR

Inspection status 28 November 2014 MR

Laboratory evaluation 09 March 2015 MR

MR: Meets Requirements NA: Not Applicable DS-EIA-HIV-AGAB-SCREEN was accepted for the WHO list of prequalified in vitro diagnostics on the basis of data submitted and publicly available information.


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Background information RPC Diagnostic Systems submitted an application for prequalification of DS-EIA-HIV-AGAB-SCREEN. Based on the established prioritization criteria, DS-EIA-HIV-AGAB-SCREEN was given priority for prequalification. Product dossier assessment RPC Diagnostic Systems submitted a product dossier for DS-EIA-HIV-AGAB-SCREEN as per the “Instructions for compilation of a product dossier” (PQDx_018 v1). The information submitted in the product dossier was reviewed by WHO staff and external experts (assessors) appointed by WHO in accordance with the internal report on the screening and assessment of a product dossier (PQDx_009 v2). Based on the product dossier screening and assessment findings, a recommendation was made to accept the product dossier for DS-EIA-HIV-AGAB-SCREEN for prequalification. Commitments for prequalification:

1. N/A

Manufacturing site inspection A comprehensive inspection was performed at the site of manufacture (22 Yablonevaya, Nizhniy Novgorod, 603093, Russia) of DS-EIA-HIV-AGAB-SCREEN in August 2014 as per the Information for manufacturers on prequalification inspection procedures for the sites of manufacture of diagnostics (PQDx_014 v1). The inspection found that the manufacturer had an acceptable quality management system and good manufacturing practices in place that ensured the consistent manufacture of a product of good quality. The manufacturer's responses to the nonconformities found at the time of the inspection were accepted 28 November 2014. Commitments for prequalification:

1. N/A Laboratory evaluation DS-EIA-HIV-AGAB-SCREEN (RPC Diagnostic Systems) is a fourth-generation immunoassay for the detection of HIV-1/2 antibodies and HIV-1 p24 antigen in human serum and plasma. A volume of 70 µl of specimen is needed to perform the assay. This type of assay requires laboratory equipment and cannot be performed in laboratories with limited facilities. Reading of the results must be performed with a spectrophotometer. In this limited performance evaluation on a panel of 1120 specimens, we found an initial sensitivity (95% CI) of 100% (99.2 - 100%) and an initial specificity (95% CI) of 98.3% (97.0 - 99.2%) compared to the reference results. The final sensitivity (95% CI) was 100% (99.2 -


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100%) and the final specificity (95% CI) was 98.8% (97.6 - 99.5%) compared to the reference results. Lot to lot variation observed was within the acceptance range. For eight seroconversion panels, DS-EIA-HIV-AGAB-SCREEN detected on average 1.25 specimens earlier than the benchmark assay (Enzygnost Anti-HIV 1/2 Plus [Siemens Healthcare Diagnostics]) and on average 0.750 specimens earlier than Vironostika HIV Ag/Ab (bioMérieux) EIA. For the mixed titer panel, DS-EIA-HIV-AGAB-SCREEN classified one specimen as false positive; all other specimens were correctly identified. For the HIV-1 p24 antigen panel, DS-EIA-HIV-AGAB-SCREEN correctly classified all specimens. For the HIV culture supernatant panel, DS-EIA-HIV-AGAB-SCREEN detected all HIV-1 subtypes, the HIV-2 culture isolate was not detected. For the 1st International Reference Panel for anti-HIV [NIBSC code 02/210], DS-EIA-HIV-AGAB-SCREEN detected all subtypes tested (HIV-1 A, HIV-1 B, HIV-C, HIV-1 CRF01_AE, HIV-1 O and HIV-2). For the HIV-1 p24 antigen standard [NIBSC code 90/636], DS-EIA-HIV-AGAB-SCREEN detected to 0.39 international units). In contrast, Vironostika HIV Ag/Ab (bioMérieux) detected to 12.5 international units. In this study, 0% of the results were recorded as indeterminate. The invalid rate was 0%.


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Labelling

1. Labels

2. Instructions for use


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1. Labels

Inside labels


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Outside labels


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2. Instructions for use


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